SUMMARYA radioimmunoassay for avian luteinizing hormone (LH) is described, using antisera raised against chicken pituitary gonadotrophins of varying degrees of purity, and purified chicken LH for radio-iodination. A postprecipitation double antibody method was developed with a sensitivity to 30-60 pg of purified chicken LH.The specificity of the method was investigated. Fractions of follicle-stimulating hormone with high biological activity showed little immunological activity, whilst all the fractions of LH tested showed strong immunological potency. Human, ovine and bovine LH showed virtually no cross-reaction.The method measures immunoreactive LH in 25-200\g=m\lplasma from chickens and quail. The plasma levels correlate well with known physiological processes, being absent in hypophysectomized birds and low in sexually immature quail. When testicular growth begins in quail the level rises eightfold; castration increases it still further while the level is lowered by testosterone. Manipulation of the pituitary-thyroid system in quail did not significantly affect the level of plasma activity.Estimates of activity are independent of either the antiserum or the iodinated preparation of LH used in the assay.The antisera used all blocked gonadotrophic activity in vivo.
Delivery of biomolecules to plants relies on Agrobacterium infection or biolistic particle delivery, the former of which is amenable only to DNA delivery. The difficulty in delivering functional biomolecules such as RNA to plant cells is due to the plant cell wall, which is absent in mammalian cells and poses the dominant physical barrier to biomolecule delivery in plants. DNA nanostructure-mediated biomolecule delivery is an effective strategy to deliver cargoes across the lipid bilayer of mammalian cells; however, nanoparticle-mediated delivery without external mechanical aid remains unexplored for biomolecule delivery across the cell wall in plants. Herein, we report a systematic assessment of different DNA nanostructures for their ability to internalize into cells of mature plants, deliver siRNAs, and effectively silence a constitutively expressed gene in Nicotiana benthamiana leaves. We show that nanostructure internalization into plant cells and corresponding gene silencing efficiency depends on the DNA nanostructure size, shape, compactness, stiffness, and location of the siRNA attachment locus on the nanostructure. We further confirm that the internalization efficiency of DNA nanostructures correlates with their respective gene silencing efficiencies but that the endogenous gene silencing pathway depends on the siRNA attachment locus. Our work establishes the feasibility of biomolecule delivery to plants with DNA nanostructures and both details the design parameters of importance for plant cell internalization and also assesses the impact of DNA nanostructure geometry for gene silencing mechanisms.
Radioimmunoassays were used to estimate luteinizing hormone (LH) and progesterone in samples of blood taken from individual hens at frequent intervals throughout their respective ovulatory cycles. A consistent pattern in the plasma levels of both hormones was observed. Significantly more LH and progesterone was present in plasma 4\p=n-\7h before ovulation than at other times during the cycle. An increase in the level of progesterone either preceded that of LH or the two hormones increased simultaneously. At no time did an increase in the level of LH occur before the rise in progesterone.In those birds which did not ovulate during the experimental period the levels of both hormones remained low. The significance of these findings in relation to the neuroendocrine control of ovulation in the hen is discussed.
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